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Speciation by postzygotic isolation: forces, genes and molecules.

New species arise as reproductive isolation evolves between diverging populations. Here we review recent work in the genetics of postzygotic reproductive isolation-the sterility and inviability of species hybrids. Over the last few years, research has taken two new directions. First, we have begun to learn a good deal about the population genetic forces driving the evolution of postzygotic isolation. It has, for instance, become increasingly clear that conflict-driven processes, like sexual selection and meiotic drive, may contribute to the evolution of hybrid sterility. Second, we have begun to learn something about the identity and molecular characteristics of the actual genes causing hybrid problems. Although molecular genetic data are limited, early findings suggest that "speciation genes" correspond to loci having normal functions within species and that these loci sometimes diverge as a consequence of evolution in gene regulation.

Animals↗

Vanadium speciation in serum by means of blue native gel electrophoresis.

Slab-gel electrophoresis has been applied to the speciation of vanadium in serum. The electrophoresis separation is an adaptation of the blue native polyacrylamide gel electrophoresis separation necessary to ensure the stability of the vanadium-protein complex; Coomassie blue was used to shift the charges of the proteins and to stabilize the vanadium complex. The detection of the vanadium species was made possible by the use of the (48)V radiotracer and the phosphor-screen technology. The method was first developed using transferrin, incubated with (48)V, as a model. After it was proved that the vanadium-transferrin complex was stable during separation, the method was validated by separating serum incubated with (48)V. The efficiency of the separation was assessed according to two parameters: resolution and conservation of the species. First, the resolution of the separation was as expected from a native separation. Second, the release of free vanadium from the transferrin complex, which was the main vanadium species expected, was negligible, which proves that the species remain intact during separation. In accordance with the literature, it was found that vanadium binds to transferrin in incubated serum at these low concentrations.

Autoradiography↗

Ti and Fe speciation by X-ray photoelectron spectroscopy(XPS) and Mössbauer spectroscopy for a full crystal chemical characterisation of Ti-garnets from Colli Albani (Italy).

Different analytical and structural methods (Electron Probe Micro-Analysis, Single Crystal X-ray diffraction, X-ray Photoelectron Spectroscopy, Mössbauer spectroscopy) were combined to fully characterise the crystal chemistry of natural Ti-bearing garnets from Colli Albani (Lazio, Italy). The study of the relevant complex crystal chemistry ( large number of cation substitutions affecting the three independent X(8-fold), Y(6-fold) and Z(4-fold) crystallographic sites and Fe and Ti transition elements exhibiting several oxidation states and coordination environments) benefited from the multi-technique approach. Electron probe microanalysis provided elemental composition of the analysed samples, which have low Ti-content (TiO2 in the range 1.99 - 3.48 wt %) and slightly different Fe/Al ratios. For all samples, two doublets were fitted to room temperature Mössbauer spectra and assigned to Fe3+(Y) (approximately 95%) and Fe2+(X) (approximately 5%). Up to three doublets (Ti3+(Y), Ti4+(Y), Ti4+(Z)) were fitted to XPS spectra and yielded direct evaluation of Ti site population. XPS technique confirmed its potential for the study of speciation of Ti in minerals.

Crystallization↗

Chromium speciation by surfactant assisted solid-phase extraction and flame atomic absorption spectrometric detection.

A new procedure has been developed for chromium speciation in aqueous solution by the use of micellar, ion-association, solid-phase extraction techniques (SPE) followed by flame atomic absorption spectrometry. The method was based on the use of C-18 bonded phase silica SPE disks for retention of ion-associated Cr(VI) with cetyl trimethyl ammonium bromide (CTAB), elution of the retained species and subsequent detection by flame atomic absorption spectrometry (FAAS). Cr(III) was oxidized by potassium persulfate to Cr(VI), then the total chromium was retained on the disk and determined by FAAS. The amount of Cr(III) was calculated by the difference between the total and Cr(VI) values. The calculated limit of detections (LOD) (based on 3sigma) are 15 microg L(-1) and 20 microg L(-1) for Cr(VI) and Cr(III) respectively. No considerable interferences have been observed from other investigated anions and cations and the method has been successfully applied to water samples taken from the Karoon River in Khuzestan province.

Chromium↗

A novel multipurpose Excel tool for equilibrium speciation based on Newton-Raphson method and on a hybrid genetic algorithm.

A new algorithm for simulation of chemical equilibria is developed, based on classical Newton-Raphson method applied to mass balance. This tool, named EST (Equilibrium Speciation Tool), is improved by using a robust Genetic Algorithm. In addition, EST works by using Excel spreadsheets and therefore offers the innovation of a great simplicity and versatility. In fact, it allows the users to simulate, or to obtain from experimental data, desired chemical-physical parameters as well as to interact with other available or freely created Excel tools. The reliability of this utility is here proved by comparison with some published data by other authors, concerning both complicated homogeneous and heterogeneous equilibria. In addiction its flexibility is tested computing thermodynamic parameters by using experimental calorimetric data referred to the complex formation of cobalt(II) with a macrocyclic ligand. A brief review and comparison of the relative robustness and quickness of main numerical methods are also reported.

Algorithms↗

Metal ion distribution between water and river sediment: speciation model and spectroscopic validation.

Natural sediments show sequestering properties that can lead to a process of self-purification of aquatic environment from metal pollution. The study of the interaction between metal ions and sediment particles enhances what is known about the distribution and bioavailability of heavy metals in natural systems. Our contribution concerns the characterisation of the sequestering ability of a River Po sediment with regard to calcium(II), magnesium(II), cadmium(II), nickel(II) and copper(II), in fixed experimental conditions, through pH-metric and spectrometric measurements. A batch titration procedure was adopted and, in each solution, after equilibration, both pH and pM (M = Ca(II), Mg(II), Cd(II), Ni(II), Cu(II)) (via Inductively Coupled Plasma - Optical Emission Spectroscopy, ICP-OES) values were measured. The experimental data were first processed with a specific software to evaluate the concentration and protonation constants of the sediment ligand site(s). The speciation model was then assessed, together with the values of complexation constants, for the different sediment/metal cation systems. In order to better characterise the copper(II)-sediment interaction and to obtain more information about the nature of ligand site(s) involved, EPR (Electronic Paramagnetic Resonance) measurements were also made on the dry sediment before and after reaction with copper(II) ions.

Biological Availability↗

Dioxouranium(VI)--carboxylate complexes. Interaction with dicarboxylic acids in aqueous solution: speciation and structure.

In this paper we report the results of an investigation performed by potentiometric (H+-glass electrode) and visible spectrophotometric measurements on the interaction of UO2(2+) ion towards some carboxylic ligands (acetate, malonate, succinate, azelate). The measurements were carried out at T= 25 degrees C in different ionic media (KNO3 and NaCl) at different ionic strengths (0.1 < or = I/mol L(-1) < or = 1.0, NaCl; I/mol L(-1) = 0.1, KNO3). The dependence on ionic strength of formation constants was taken into account by using both a simple Debye-Hückel type equation and the SIT (Specific ion Interaction Theory) approach. Different speciation models (depending on concentration of reagents, ionic strength, pH-range) both for different carboxylates and different ionic media have been obtained. Linear combinations between formation constants, stoichiometric coefficients and length of alkyl chain of dicarboxylates have been observed and predicted formation constants at I= 0 mol L(-1) are reported for the interaction of UO2(2+) with HOOC-(CH2)n-COOH with 1 < or = n < or = 7. Finally, a visible absorption spectrum for each complex reaching a significant percentage of formation in solution (KNO3 medium) has been calculated to characterise the compounds found by pH-metric refinement.

Dicarboxylic Acids↗

Speciation and inversions: chimps and humans.

A new set of models has resurrected a role for chromosomal inversions in the formation of new species. Traditional models, which are generally considered to be unlikely in most cases, had imagined that inversions might aid speciation by directly causing low hybrid fitness. In contrast, the newer models focus on the effect that inversions have on local recombination rates. A test of these models found a strikingly high rate of amino-acid substitution within regions where humans and chimpanzees differ by inversions, suggesting perhaps that our ancestral species underwent a divergence process in which gene flow and inversions played a key role. However, it remains uncertain whether this interesting finding is actually consistent with the proposed model.

Animals↗

Effect of yeast extract on speciation and bioavailability of nickel and cobalt in anaerobic bioreactors.

The speciation of metals plays an important role in their bioavailability. In the case of anaerobic reactors for the treatment of wastewaters, the ubiquitous presence of sulfide leads to extensive precipitation of metals like nickel and cobalt, which are essential for the metabolism of the anaerobic microorganisms that carry out the mineralization of the pollutants present in the wastewater. In practice, nickel, cobalt, and iron are added in excessive amounts to full-scale installations. This study is concerned with the complexation of nickel and cobalt with yeast extract and its effect on the biogas production by methanogenic biomass. Adsorptive stripping voltammetry (AdSV) was used to get information about the stability and complexing capacity of the metal-yeast extract complexes formed. Nickel and cobalt form relatively strong organic complexes with yeast extract. The bioavailability of these essential metals in anaerobic batch reactors was dramatically increased by the addition of yeast extract. This is due to the formation of dissolved bioavailable complexes, which favors the dissolution of metals from their sulfides. Trace doses of yeast extract may be effective in keeping additions of essential metals to anaerobic reactors at a minimum.

Anaerobiosis↗

Hybridation of different chiral separation techniques with ICP-MS detection for the separation and determination of selenomethionine enantiomers: chiral speciation of selenized yeast.

Enantioseparation and determination of selenomethionine enantiomers in selenized yeast was investigated using chiral separation techniques based on different principles, coupled on-line to inductively coupled plasma mass spectrometry (ICP-MS) for selenium-specific detection. High performance liquid chromatography (HPLC) on a beta-cyclodestrin (beta-CD) column, cyclodextrin-modified micellar electrokinetic chromatography (CD-MEKC), gas chromatography (GC) on a Chirasil-L-Val column, and HPLC on a Chirobiotic T column have been investigated as the chiral separation techniques. For HPLC separation on the beta-CD column, and also for CD-MEKC, selenomethionine enantiomers were derivatized with NDA/CN(-). For chiral separation by GC, selenomethionine enantiomers were converted into their N-trifluoroacetyl (TFA)-O-alkyl esters. The developed hybridation methodologies are compared with respect to enantioselectivity, sensitivity and analysis time. The usefulness of the best-suited method [HPLC (Chirobiotic T)-ICP-MS] was demonstrated by its application to the successful chiral speciation of selenium and D-and L-selenomethionine content determination in selenized yeast.

Chromatography, Gas↗

Fast speciation analysis of iodophenol compounds in river waters by capillary electrophoresis-inductively coupled plasma-mass spectrometry with off-line solid-phase microextraction.

An analytical methodology for the fast separation and determination of iodophenol species in natural water samples was developed using capillary electrophoresis (CE) coupled to inductively coupled plasma-mass spectrometry (ICP-MS). Based on the element-specific and highly sensitive detection provided by ICP-MS, the methodology has been applied to the analysis of 2-iodophenol, 4-iodophenol, and 2,4,6-triiodophenol. The use of solid-phase microextraction (SPME), after proper optimization, improved the signal by a factor of 100 leading to detection limits in the sub microg.L(-1). Different desorption conditions of iodophenol compounds from the SPME microfiber were studied to achieve the optimum preconcentration factor and best analytical performance. Different CE conditions were studied to achieve complete baseline separation of iodophenols in short migration times. Three different CE buffer systems were evaluated using ICP-MS detection. A buffer solution containing 20 mmol.L(-1) 3-(cyclohexylamino)-1-propanesulfonic acid (CAPS) and an applied potential of +22 kV were finally selected leading to a maximum separation time of 6.6 min. A relative standard deviation (%RSD) of about 5.0% for ten consecutive determinations was obtained. Finally, the speciation methodology developed was utilized for the determination of iodophenol compounds in natural water samples.

Buffers↗

Development of a new hybrid technique for rapid speciation analysis by directly interfacing a microfluidic chip-based capillary electrophoresis system to atomic fluorescence spectrometry.

This paper represents the first study on direct interfacing of microfluidic chip-based capillary electrophoresis (chip-CE) to a sensitive and selective detector, atomic fluorescence spectrometry (AFS) for rapid speciation analysis. A volatile species generation technique was employed to convert the analytes from the chip-CE effluent into their respective volatile species. To facilitate the chip-CE effluent delivery and to provide the necessary medium for subsequent volatile species generation, diluted HCl solution was introduced on the chip as the makeup solution. The chip-CE-AFS interface was constructed on the basis of a concentric "tube-in-tube" design for introducing a KBH4 solution around the chip effluent as sheath flow and reductant for volatile species generation as well. The generated volatile species resulting from the reaction of the chip-CE effluent and the sheath flow were separated from the reaction mixture in a gas-liquid separator and swept into the AFS atomizer by an argon flow for AFS determination. Inorganic mercury (Hg(II)) and methylmercury (MeHg(I)) were chosen as the targets to demonstrate the performance of the present technique. Both mercury species were separated as their cysteine complexes within 64 s. The precision (relative standard deviation, RSD, n = 5) of migration time, peak area, and peak height for 2 mg.L(-1) Hg(II) and 4 mg.L(-1) MeHg(I) (as Hg) ranged from 0.7 to 0.9%, 2.1 to 2.9%, and 1.5 to 1.8%, respectively. The detection limit was 53 and 161 microg.L(-1) (as Hg) for Hg(II) and MeHg(I), respectively. The recoveries of the spikes of mercury species in four locally collected water samples ranged from 92 to 108%.

Electrophoresis, Microchip↗

Mercury speciation by CE: a review.

CE methods for the speciation of inorganic and organomercury compounds are reviewed. Sample preparation, separation conditions and detection modes are discussed. Efficient separation and sensitive determination of mercury species by CE typically involves complexation with various thiols, chromogenic and other chelating agents; however, some methods do not require complexation. Spectrophotometric detection based on UV-visible absorption is by far the most commonly used. Hyphenated techniques, such as CE/inductively coupled plasma (ICP)-MS, hydride generation coupled to ICP-MS or atomic fluorescence spectrometry and CE/atomic absorption spectrometry are gaining popularity due to their high sensitivity and selectivity. Last, but not least, the potential and applications of electrochemical methods for detection of separated mercury species are outlined.

Electrochemistry↗

The toxicology of chromium with respect to its chemical speciation: a review.

The properties of trivalent and hexavalent chromium are reviewed with respect to acute and chronic oral toxicity, dermal toxicity, systemic toxicity, toxicokinetics, cytotoxicity, genotoxicity and carcinogenicity. The hexavalent chromium compounds appear to be 10-100 times more toxic than the trivalent chromium compounds when both are administered by the oral route. Dermal irritancy and allergy are more frequently caused by contact with soluble hexavalent chromium compounds. The cytotoxicity of soluble and insoluble hexavalent chromium compounds to fibroblasts is 100-1000 times greater than that demonstrated by trivalent chromium compounds. In short-term tests, the hexavalent chromium compounds demonstrated genotoxic effects four times more frequently than did the trivalent chromium compounds. Carcinogenicity appears to be associated with the inhalation of the less soluble/insoluble hexavalent chromium compounds. The toxicology of chromium does not reside with the elemental form. It varies greatly among a wide variety of very different chromium compounds. Oxidation state and solubility are particularly important factors in considering the toxicity of chromium with respect to its chemical speciation.

Animals↗

Cadmium uptake in rat hepatocytes in relation to speciation and to complexation with metallothionein and albumin.

Cadmium (Cd) uptake has been studied in primary cultures of rat hepatocytes focusing on the impact of inorganic and organic speciation. Uptake time-course studies over a 60-min exposure to 0.3 microM (109)Cd revealed a zero-time uptake and a slower process of accumulation which proceeds within minutes. (109)Cd uptake showed saturation kinetics (K(m) = 3.5 +/- 0.8 microM), and was highly sensitive to inhibition by Zn and Hg. There was no evidence for sensitivity to the external pH nor for any preferential transport of the free cation Cd(2+) over CdCl(n) (2-n) chloro-complexes. According to the assumption that only inorganic metal species are available, metal uptake decreased upon albumin (BSA) addition to the exposure media. In contrast, higher levels of (109)Cd accumulation were obtained under optimal conditions for Cd complexation by MT. Comparison among uptake data obtained under inorganic and organic conditions revealed that Cd-MT would be taken up 0.4 times as rapidly as Cd(inorg). We conclude that uptake of Cd in rat hepatocytes involves specific transport mechanism(s) subjected to Zn or Hg interactions. Uptake of inorganic Cd is not proportional to the levels of free Cd(2+) and does not involve the divalent cation transporter DCT1 nor the co-transporter Fe(2+)-H(+) NRAMP2. We found Cd-MT but not Cd-BSA to be available for the liver cells, and have estimated a binding affinity four orders of magnitude higher for Cd complexation with MT compared to BSA; MT may have a significant role in Cd delivery to the liver.

Albumins↗

Serological speciation of human T-cell leukaemia virus infections using synthetic peptide antigens.

In order to assess the specificity and sensitivity of two peptide-based assays (Synth HTLV-I and HTLV-II enzyme-linked immunoassay [EIA] [UBI] and Select-HTLV EIA [IAF]) in discriminating between antibody to HTLV-I and HTLV-II infection, a panel of 186 well-characterised serum/plasma samples was tested by the two assays. The panel comprised 160 samples that by Western blot were confirmed to contain antibodies to HTLV-I/II and 26 samples that showed reactivity with gag but not env gene products. Both assays were found to be specific in that they did not misclassify any of the 80 specimens from cases of tropical spastic paraparesis or adult T-cell leukaemia/lymphoma, diseases believed to be HTLV-I associated, as anti-HTLV-II positive. Of the 160 specimens confirmed as anti-HTLV-I/II positive by Western blot, 6.2% were negative or untypable in the Synth EIA compared with 13.7% in the Select EIA. Of the 26 Western blot indeterminate samples, 16 were negative by both assays. Five were typed as anti-HTLV-I by both assays and 5 as anti HTLV-II by Select EIA only. The peptide based EIAs offer an economical and, in most cases, reliable means of discriminating between anti-HTLV-I and anti-HTLV-II. However, they should only be applied to sera that have been confirmed by Western blot or other methods as anti-HTLV-I/II positive. Even then they may fail to speciate sera from non-Japanese, non-Afrocaribbean populations.

Blotting, Western↗

On-line separation for the speciation of mercury in natural waters by flow injection-cold vapour-atomic absorption spectrometry.

Inorganic mercury and methylmercury are determined in natural waters by injecting the filtered samples onto a low cost commercial flow injection system in which an anion exchange microcolumn is inserted after the injection loop (FIA-IE). If hydrochloric acid is used as the carrier solution, the HgCl4(2-) species (inorganic mercury) will be retained by the anion exchanger while the CH3HgCI species (methylmercury) will flow through the resin with negligible retention. Four anion exchangers and seven elution agents were checked, in a batch mode, to search for the best conditions for optimal separation and elution of both species. Dowex M-41 and L-cysteine were finally selected. Mercury detection was performed by cold vapour-electrothermal atomic adsorption spectrometry (HG-ETAAS). Both systems were coupled to perform the continuous on-line separation/detection of both inorganic mercury and methylmercury species. Separation and detection conditions were optimized by two chemometric approaches: full factorial design and central composite design. A limit of detection of 0.4 microg L(-1) was obtained for both mercury species (RSD < 3.0% for 20 microg L(-1) inorganic and methylmercury solutions). The method was applied to mercury speciation in natural waters of the Nerbioi-lbaizabal estuary (Bilbao, North of Spain) and recoveries of more than 95% were obtained.

Animals↗

On-line speciation and determination of Cr(III) and Cr(VI) in drinking and waste water samples by reversed-phase high performance liquid chromatography coupled with atomic absorption spectrometry.

A simple, rapid, and selective on-line method for the speciation and determination of Cr(III) and Cr(VI) in aqueous solutions by ion-pairing HPLC coupled with flame atomic absorption spectrometry (FAAS) is described. The composition of the mobile phase has been optimized for better separation. The effects of column temperature, volume of injection loop, fuel flow rate of FAAS, and nebulizer suction rate of FAAS have also been investigated. Separation is accomplished in almost 2.5 min on a 25 cm length C18 column at 40 degrees C. The selectivity of the method has been established by investigating the effect of interfering elements on chromium determination. The detection limit (3sigma) achieved by the method was calculated as 3.7 ng/mL for Cr(III) and 2.0 ng/mL for Cr(VI). The proposed method has been validated by analyzing certified reference material (BCR 544) and successfully applied to the analysis of drinking water and wastewater samples with a relative error below 6%.

Journal Article↗